Basic Vector Information
- Vector Name:
- pLenti-Puro-sgAMPKa2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13013 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- U6
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLenti-Puro-sgAMPKa2 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pLenti-Puro-sgAMPKa2 vector Sequence
LOCUS 62056_13875 13013 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13013)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..13013
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..199
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 444..569
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1062..1295
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1480..1524
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 1673..1714
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 1822..1939
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1990..2230
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 2260..2335
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 2397..2608
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 2633..6736
/label=Cas9
/note="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
CDS 6785..6808
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 6818..6874
/label=P2A
/note="2A peptide from porcine teschovirus-1 polyprotein"
CDS 6875..7468
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 7487..8075
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 8147..8380
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 8412..8636
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 8682..9110
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 9124..9453
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 9501..9548
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 9567..9938
/label=BleoR
/note="antibiotic-binding protein"
promoter 10000..10202
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter complement(10289..10319)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(10334..10355)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(10643..11231)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11405..12262)
/label=AmpR
/note="beta-lactamase"
promoter complement(12263..12367)
/label=AmpR promoter
enhancer 12633..13012
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
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